Objective:To investigate the effect of dimethyl amiloride (DMA) on invasive activity of PGCL3 cells from a human highly-metastatic lung carcinoma cell line in vitro and elucidate its possible mechanism. Methods:The invasion and migration capacities of PGCL3 cells were measured by using Transwell chamber assay after pretreatment with DMA. The effects of DMA on the activity of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) secreted by PGCL3 cells were measured by chromogenic substrate assay. The effects of DMA on uPA, urokinase-type plasminogen activator receptor (uPAR), and PAI-1 mRNAs transcription were determined by RT-PCR. The expression levels of extracellular regulated protein kinases 2 (ERK2) and ras protein were assessed by Western blot. Results:DMA inhibited invasion and migration capabilities of PGCL3 cells in vitro, down-regulated the mRNA transcription of uPA, uPAR and PAI-1, as well as up-regulated the expression of ras protein. After 24-hour treatment, DMA reduced the activity of uPA at higher concentration, but DMA had no effects on the activity of secreted PAI-1 protein and expression of ERK2 protein. Conclusion:DMA inhibits the invasion and migration of highly-metastatic lung cancer PGCL3 cells. The mechanism might be associated with down-regulation of the expression of uPA system.

Sleep disorders are common diseases with various dys-function during sleep-wake process, including difficulty falling or staying asleep, falling asleep at inappropriate times, excessive total sleep time, or abnormal behaviors associated with sleep. Sleep disorders can lead to the deposition of amyloid beta protein ( Aβ) by affecting the normal metabolism of amyloid beta protein in the brain. Patients with Alzheimer’s disease ( AD) often suffer from sleep disorders, and its pathology always results in more se-vere sleep disorders, which leads to a risk of cognitive impair-ment and hypofunction. Sleep disorders could interact closely with AD, forming a positive feedback loop, which causes serious damage to the body health. This review summarized the current research about sleep disorder in the onset of AD and the current status of medication.

Objective: To investigate the effect of amiloride on in vitro invasive activity and uPA (urokinase-type plasminogen activator)system of human highly metastatic lung carcinoma cell line PGCL3. Methods: At 6 hours after treatment with amiloride at the concentrations of 25μmol/L，50μmol/L and 100μmol/L for PGCL3 cells，Transwell Chamber assay was performed to detect the effect of amiloride on the invasive and migratory capacity of PGCL3 cells.Effect of amiloride on the activity of uPA and PAI-1(plasminogen activator inhibitor-1)secreted by PGCL3 cells were measured by chromogenic substrate assay after PGCL3 cells were incubated with amiloride for 24 hours.RT-PCR was used to analyze the effect of amilorede on mRNA levels of uPA，uPAR(urokinase-type plasminogen activator receptor)and PAI-1.The expression levels of uPA，ERK2(extracellular regulated protein kinases 2)and ras protein were assessed by Western blot. Results: The number of cells through membrane was significantly decreased in invasion and migration test in vitro.The inhibitory rates of invasion and migration after treatment with amiloride of 100μmol/L were 37.7%±4.1%and 64.9%±4.9%.respectively,with a significant difference from those in the control group(P<0.01).At 24 hours after amiloride treatment，the chromogenic substrate assay showed direct inhibition of the activity of uPA and PAI-1 secreted by PGCL3 cells.No effect on the expression of uPAR in mRNA level was observed,but the expression of PAI-1 in mRNA level was significantly inhibited.Amiloride of 100μmol/L dramatically inhibited the expression of uPA mRNA.The expression level of uPA protein was decreased with the increase of the concentration of amiloride,but no effect was observed on the expression of ERK2 and ras in protein level.Conclusion: Amiloride can inhibit the invasion and migration of PGCL3 cells,through inhibiting the expression and activity of uPA and PAI-1.Amiloride is a potential agent to inhibit cancer invasion and metastasis.

Aim To establish a hepatic carcinoma cell line with stable voltage-gated chloride channel 3 ( ClC-3 ) gene silencing through the lentivirus-mediated short-hairpin RNA ( shRNA ) method and investigate the effects of gene silencing on invasion and migration. Methods Three lentiviral vectors coding shRNA tar-geting ClC-3 gene were constructed, the recombinant plasmids were packaged into mature lentivirus by 293FT cells, and then the lentiviruses were harvested, concentrated and titrated. MHCC97H cells were infec-ted with the recombinant lentiviruses and then were se-lected to obtain cell lines stably expressing ClC-3 shR-NA. The efficiency of ClC-3 mRNA and protein ex-pression interference were determined by real-time flu-orescence quantitative PCR and Western blot, respec-tively. The effects of ClC-3 gene interference on inva-sion and migration of MHCC97 H cells were performed by Transwell chamber assays with or without Matrigel and cell scratch assay. Results The recombinant lentiviral vectors were successfully constructed and four lentiviruses were acquired after packaged by 293 FT cells. One negative control cell line and three cell lines with ClC-3 gene interference ( MHCC97 H/shClC-3-1 , shClC-3-2 and shClC-3-3 ) were successfully construc-ted after MHCC97 H cells were infected with lentivirus-es. The expression level of ClC-3 mRNA and protein in three ClC-3-silenced cells were obviously lower than the negative control cells ( P <0. 01 ) , MHCC97 H/shClC-3-2 cells showed the greatest inhibition of ClC-3 mRNA and protein expressions. As compared with the negative control cells, the ClC-3 gene interference sig-nificantly decreased invasion and migration of MH-CC97 H cells in vitro ( P <0. 01 ) . Conclusion The hepatic carcinoma cell lines with stable ClC-3 gene si-lencing were successfully established and the ClC-3 gene interference could significantly inhibit invasion and migration of MHCC97H cells.

AIM: To investigate the expression of stem cell factor (SCF) in tryptase -positive mast cells ( MCs) in different types of human periapical diseases for determining the role of SCF and MCs in the pathogenesis of peria-pical diseases.METHODS: A total 50 cases of specimens were involved in this study, including healthy control (n=20), periapical cyst (n=15) and periapical granuloma (n=15).The tissue material was fixed in 10%formalin for at least 48 h, stained with hematoxylin and eosin for the observation of histopathology, stained with immunohistochemistry for identifying MCs and MCs degranulation, and stained with double immunofluorescence for identification of tryptase-SCF double positive MCs.RESULTS:Compared with healthy control, significantly higher densities of both total and degranu-lated MCs in human periapical lesions were observed.The densities of both total and degranulated MCs in the periapical cyst were significantly higher than that in the periapical granuloma.The density of tryptase-SCF double positive MCs in the periapical lesions was significantly higher than that in the healthy controls.The density oftryptase-SCF double positive MCs in the periapical cyst was significantly higher than that in periapical granuloma.No significant difference in the density of MCs between immunohistochemistry staining and double immunofluorescence staining was observed.CONCLUSION:The tryptase-SCF double positive MCs play an active role in the pathogenesis of the periapical inflammatory lesions, particularly in the formation of fibrous tissue in periapical cyst.The potential role of the tryptase-SCF double positive MCs relates with the initiation, development, and persistence of the periapical inflammatory process.

This study is to investigate the sedative and hypnotic effects of a novel compound H1208. The sedative activity of H1208 was investigated by recording the spontaneous locomotor activity of mice. The hypnotic property was evaluated by the latency and duration of sleep (loss of righting reflex) in mice and the effect of hypnotics on sleep pattern of electroencephalogram were studied in conscious, freely moving mice with chronically implanted electrodes. The brain monoamine neurotransmitters levels in mice were measured by high performance liquid chromatography-electrochemical detection. The spontaneous locomotor activity was decreased by 56.7% and 80.2% in H1208 (5 and 25 mg x kg(-1), ip) treated mice, respectively. The loss of righting reflex was directly induced in mice after H1208 (60 mg x kg(-1), ip) administration. The non-rapid eye movement sleep increased significantly by 131% and 259%, respectively, within 3 hours after H1208 (30 and 60 mg x kg(-1), ip) administration. However, the rapid eye movement sleep decreased significantly. The contents of DA in the striatum and cortex and 5-HT in the cortex decreased significantly. These results demonstrated that H1208 has potent sedative and hypnotic effects, which may be closely related to the decreased contents of DA and 5-HT in mouse brain.

A series of isoindoline derivatives were designed, synthesized, and evaluated for their double inhibitory activities. All of them were new compounds, and their structures were confirmed by 1H NMR and HR-MS. Preliminary in vitro pharmacological tests showed that all compounds exhibited 5-HT or NE reuptake inhibition activity. Among the tested compounds, compound I-3 exhibited potent inhibitory activity against 5-HT and NE reuptake in vitro, and exhibited potent antidepressant activity in vivo. These compounds designed can be further optimized for finding more potent 5-HT/NE dual reuptake inhibitors and antidepressant candidates as well.

Objective To evaluate two treatment schemes of liver protection and choose the optimal radiotherapy using phar‐macoeconomics principle and methods .Methods In‐patients who were diagnosed as decompensated cirrhosis patients and simultaneously conformed to the Chlid‐Pugh graduation of level B‐C during 2010-2012 were chosen .The chosen patients were divided into A ,B and C group separately ,and were treated with the compound prescription of dichloro acetic acid two isopropyl amine inoculation fluid ,the poly‐ene phosphatide acid radical choline inoculation fluid ,and the compound prescription dichloro acetic acid two isopropyl amine inoculation fluid and the polyene phosphatide acid radical choline inoculation fluid .Cost effectiveness analysis was done between group B and C .Re‐sults Treatment time of group B and C was (15 .5 ± 8 .55) d and (12 .13 ± 7 .61) d respectively .The total effectiveness was 50% and 60% respectively and the difference was with non‐statistics significance (P>0 .05) .The total cost was (1 640 .52 ± 905 .26) yuan and (2 576 .51 ± 1 615 .99) yuan respectively .Conclusion Group B was the better treatment scheme .

Objective To find out the mechanism of strephenopodia and strephexopodia of children with spastic cerebral palsy by analyzing their gait character.MethodsForty children with spastic cerebral palsy and forty healthy children with normal walking ability were involved in this research.Footscan 7 gait analysis system was used to measure foot initial touchdown part,the ratio of different part touchdown phase to single supporting phase,the degree of strephenopodia and strephexopodia in different part.Two groups' characteristic parameters were analyzed by statistics method.ResultsSignificant differences were found in foot first touchdown part between two groups(P<0.05).The phenomenon that toes or metatarsus or whole foot first touch the earth was found in children with spastic cerebral palsy.The ratio of anterior foot and middle foot touchdown phase to single supporting phase were higher than normal children(P<0.05).In spastic cerebral palsy group the phenomenon of strephexopodia was more serious than strephenopodia during anterior foot touchdown phase.ConclusionLots of children with spastic cerebral palsy are suffering long-time compression pain and strephexopodia with anterior foot,active rectification must be performed during rehabilitation care.

Skin sensitization (allergic contact dermatitis, ACD), is a serious condition caused by small reactive molecules and is characterized by a delayed-type hypersensitivity .Animal tests were usually used in the evaluation of sensitizing potential of chemical substances in the past .However , there is an increasing interest from the public for reducing and ultimately replacing animal tests .The European Union (EU) has posed a ban on animal testing of cosmetic ingredients that includes skin sensitization since 2013.Therefore, alternative in vitro tests are the main tendency in chemical substances and cosmetic sensitizing potential research in the future .In this study, different kinds of in vitro test methods that were adopted by OECD or on research (LLNA, DPRA, KeratinoSens TM, h-CLAT) were reviewed through recent years literature , comprehensive introduction and evaluation were made to obtain reliable hazard and potency information on potential skin sensitizers .